Discovery and verification of methylated circulating tumor DNA markers for the detection of colorectal cancer in subjects under 50 years of age

发现并验证用于检测 50 岁以下受试者结直肠癌的甲基化循环肿瘤 DNA 标记

• 批准号: 10306165
• 负责人: ROBERT William HAILE
• 金额: $ 68.55万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10306165
• 关键词: Address; Adherence; Adult; Advisory Committees; Age; Age of Onset; Age-Years; Biological Assay; Biological Markers; Blood; Case-Control Studies; Characteristics; Colon; Colonoscopy; Colorectal; Colorectal Cancer; DNA; DNA Markers; DNA methylation profiling; Data; Data Set; Detection; Diagnosis; Diagnostic; Disease; Early Diagnosis; Ensure; FDA approved; Feces; Formalin; Freezing; Gold; Human; Incidence; Leukocytes; Life; Malignant Neoplasms; Methods; Methylation; Mucous Membrane; Mutation; Neoplasms; Paraffin Embedding; Pathway interactions; Patients; Performance; Persons; Plasma; Population; Preventive service; Primary Neoplasm; Research Design; Resistance; Risk; SEER Program; Sampling; Screening for cancer; Series; Signal Transduction; Site; Specificity; Symptoms; Testing; The Cancer Genome Atlas; Time; Tissues; Triage; Tumor stage; Validation; adenoma; advanced disease; age group; aged; base; bioinformatics pipeline; biomarker panel; cancer type; case control; colon cancer patients; colorectal cancer screening; colorectal cancer treatment; cost effective; diagnostic accuracy; early onset; early onset colorectal cancer; epigenomics; follow-up; head-to-head comparison; high risk; improved; metastatic colorectal; minimally invasive; mortality; novel; nuclease; population based; pyrosequencing; screening; screening guidelines; specific biomarkers; surveillance data; tumor; tumor DNA; uptake

Abstract In recent years, the incidence of colorectal cancer (CRC) under the age of 50 years has increased significantly and population-based screening is currently not offered to persons under 50 years. Consequently, persons with early-onset (<50 years) CRC (EOCRC) more frequently present with symptomatic disease at advanced stages (III/IV) resulting in greater loss of life in young cases. In 2020, the U.S. Preventive Services Task Force recommended the age for CRC screening by colonoscopy or fecal tests be reduced to 45 years. However, the uptake of screening by these methods in all screen-eligible populations is low, including in those <50 years who are genetically at high risk, so adherence in the asymptomatic population <50 years is also likely to be low. Also, over half of EOCRC occur in persons under <45 years of age, where no such screening would be offered. A minimally invasive, blood-based screening test for EOCRC would provide a cost-effective and patient-friendly option for triaging and identifying those warranting a follow-up colonoscopy, while increasing screening adherence in the younger population. Cancer-specific methylated DNA biomarkers are highly suited for population-based cancer screening via the detection of circulating tumor DNA (ctDNA) in blood plasma because they are more prevalent across patients with a given cancer type than tumor mutations and are more stable (nuclease-resistant) in plasma. The methylated SEPTIN9 plasma ctDNA test, “mSEPT9” (Epi proColon V2.0, Epigenomics), is FDA-approved for CRC screening in persons aged ≥50 years who decline colonoscopy and fecal tests. This test urgently needs to be assessed in persons <50 years to determine its suitability for the detection of EOCRC. Even so, a multi-marker test is likely to have superior sensitivity to a single-marker test. Thus, the objectives of our study are to identify and confirm a panel of methylated ctDNA markers for the plasma-based detection of EOCRC, and to compare the diagnostic performance of this panel to the mSEPT9 test in persons <50 years of age. In Aim 1, we will identify the most prevalent differentially-methylated (tumor vs. normal) DNA markers in CRC cases <50 years by performing deep Methyl-Seq across 4.2 million CpG sites in paired primary tumor vs. normal colon tissues from EOCRC cases, and leukocytes from healthy controls (to filter out non-specific markers). The identified markers will be validated in independent sample series. For the top-ranked markers, we will then develop methylation-specific real-time PCR assays with high analytical sensitivity and test these in pooled plasma from metastatic CRC cases (to select markers producing the strongest signals) and healthy controls (to eliminate any producing low-level non-specific signals) to finalize the marker panel. In Aim 2, we will evaluate the diagnostic performance characteristics of the mSEPT9 test vs. the multi-marker panel test in plasma from colonoscopy-verified CRC cases and controls <50 years. This study will yield a multi-marker methylated ctDNA panel with improved diagnostic performance over the single-marker mSEPT9 test for cost-effective, blood-based, CRC screening in asymptomatic persons <50 years of age.

抽象的 近年来，大肠癌的事件（CRC）在50岁以下已显着增加 目前没有向50岁以下的人提供基于人群的筛查。因此，人 早发（<50年）CRC（EOCRC）在晚期症状性疾病中更频繁地出现 阶段（iii/iv）导致年轻病例的生命损失更大。 2020年，美国预防服务工作队 建议通过结肠镜检查或粪便测试的CRC筛查年龄减少到45年。但是， 这些方法在所有符合屏幕资格的人群中的筛查吸收较低，包括在50年中 通常处于高风险的人，因此无症状人口<50年的依从性也可能是 低的。同样，超过一半的eocrc发生在<45岁以下的人中，没有这样的筛查 提出。对EOCRC的微创，基于血液的筛查测试将提供成本效益，并且 对患者友好的选择，用于分类和识别警告后续结肠镜检查的选择，同时增加 筛查年轻人口的依从性。癌症特异性甲基化的DNA生物标志物非常适合 用于基于人群的癌症筛查，通过检测血浆中循环肿瘤DNA（CTDNA） 因为它们在给定癌症类型的患者中比肿瘤突变更普遍，并且更多 血浆中的稳定（耐核酸酶）。甲基化的Septin9等离子体CTDNA测试，“ MSEPT9”（Epi Procolon v2.0，表观基因组学）是FDA批准的，用于降低结肠镜检查的患者的CRC筛查 和粪便测试。迫切需要对该测试进行<50年的人进行评估，以确定其适用性 检测Eocrc。即便如此，多标记测试可能对单标记测试具有较高的敏感性。 那就是我们研究的目标是识别和确认一组甲基化的ctDNA标记 基于等离子体的EOCRC检测，并将该面板的诊断性能与MSEPT9进行比较 以<50岁的人的身份进行测试。在AIM 1中，我们将确定最普遍的不同甲基化（肿瘤） 在CRC病例中，通过在420万CPG上进行深甲基序列，在CRC病例中与正常）DNA标记 配对的原发性肿瘤与EOCRC病例的正常结肠组织中的位点，以及健康的白细胞 控件（滤除非特异性标记）。确定的标记将在独立样本中进行验证 系列。对于排名最高的标记，我们将开发高甲基化特定的实时PCR测定 分析灵敏度并在转移性CRC病例的汇集血浆中测试这些敏感性（选择产生标记物 强信号）和健康控制（消除任何生产的低级非特异性信号）以最终确定 标记面板。在AIM 2中，我们将评估MSEPT9测试V的诊断性能特征。 来自结肠镜检查验证的CRC病例和对照<50年的血浆中的多标记面板测试。这项研究 将产生一个多标记的甲基化ctDNA面板，并在单标志物上提高诊断性能 MSEPT9测试在不对称的人中进行成本效益，基于血液的CRC筛查。